High-Speed Two Dimensional Separations Using Micro Free Flow Electrophoresis

在化学系化学测量和成像项目的支持下，明尼苏达大学双城分校的Michael Bowser教授和他的团队将使用微自由流动电泳（uFFE）作为二维分离的第二阶段。FFE是一种连续分析技术，能够在第一个分离柱洗脱时进一步分离分析物峰。连续分析消除了在以前的2D分离中遇到的由采样不足引起的限制。耦合毛细管电泳（CE）或纳米液相色谱（nLC）预计是直接的，没有复杂的阀门或定时需要。预计与uFFE的耦合将使这些技术的峰值容量增加20到30倍，而不会牺牲第一维分离的效率或分离时间。在15分钟的分离时间内，总2D峰值容量预计将达到2,000。计划进行实验，以充分表征使用这种方法可能的许多分离模式组合的峰值容量和兼容性。作为高速2D分离的潜在力量的演示，CE-uFFE将被纳入在线微透析分析中，用于测量体内神经化学动力学。在线微透析需要复杂混合物的高速分离，使该分析成为展示本提案中开发的2D分离优势的理想应用。该方法的技术优势在于在不牺牲分离时间的情况下提高了峰值容量。因此，二维uFFE具有广泛采用的潜力，可以提高许多研究领域的研究能力和效率。除了实际优势之外，探索2D-uFFE将为分离科学的基础提供独特的见解。密歇根大学双子城的研究人员还打算设计本提案中描述的研究项目，目标是通过丰富知识、指导和培训机会、包容代表性不足的群体和增强基础设施，最大限度地发挥其对社会的影响。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Michael Bowser at University of Minnesota-Twin Cities and his group will use micro free flow electrophoresis (uFFE) as the second stage in 2D separations. FFE is a continuous analysis technique capable of further separating analyte peaks as they elute off the first separation column. Continuous analysis removes limitations encountered in previous 2D separations caused by under sampling. Coupling with capillary electrophoresis (CE) or nano liquid chromatography (nLC) is expected to be straightforward with no complicated valving or timing required. It is anticipated that coupling with uFFE will increase the peak capacity of these techniques 20 to 30-fold with no sacrifice of efficiency or separation time in the first dimension separation. Total 2D peak capacities are expected to reach 2,000 in separation times of 15 min. Experiments are planned to fully characterize the peak capacity and compatibility of the many combinations of separation modes possible using this approach. As a demonstration of the potential power of high speed 2D separations CE-uFFE will be incorporated into online microdialysis assays for measuring in vivo neurochemical dynamics. Online microdialysis requires high-speed separations of complex mixtures, making this assay an ideal application to demonstrate the advantages of the 2D separations developed in this proposal. The technical benefit of the proposed approach lies in its increased peak capacity without sacrificing separation time. 2D uFFE therefore has the potential to be adopted widely, improving research capabilities and efficiency in many research areas. Beyond practical advantages, exploration into 2D-uFFE will provide unique insight into the fundamentals of separation science. The researchers at UM-twin cities also intend to design the research project described in this proposal with the goal of maximizing its impact on society through intellectual enrichment, mentoring and training opportunities, inclusion of underrepresented groups and infrastructure enhancement.